Phytase enzyme is used as a dietary supplement in broiler nutrition to improve phosphorous bioavailability. Phytase deliberates phosphate groups from phytic acid and produces myo-inositol after total dephosphorylation. Myo-inositol is a bioactive compound having beneficial modulatory effects on metabolism in humans. However, it is not well understood if and how phytic acid degradation products, particularly myo-inositol, can modulate metabolism in broiler chicken. The purpose of this study was to investigate effects of dietary supplements of phytase and myo-inositol on the blood plasma metabolome profile of broiler chickens. Broilers were provided a nutrient-adequate control diet or the same diet supplemented with either 3.5 g myo-inositol or 500, 1500 or 3000 units of phytase, per kilogram of feed (grower diet). Broilers were group-housed in floor pens (eight pens per diet) and provided one of the treatment diets for 22 days. Then, blood was collected from one bird per pen, resulting in eight replicated measurements per diet. A targeted metabolomics approach was applied to the heparin plasma. Body weight of the birds was not significantly affected by the treatments. Plasma myo-inositol concentrations were significantly increased by myo-inositol supplementation and phytase supplementation at 500 and 1500 units/kg. Metabolites generally affected by phytase supplementation belonged to the groups of acyl-carnitines, phosphatidylcholines, sphingomyelins, lysophosphatidylcholine, biogenic amines and amino acids. Compared to the control diet, phytase supplements had significantly higher plasma concentrations of kynurenine and creatinine, but lower concentrations of histamine and cis-4-hydroxyproline. Myo-inositol supplementation significantly increased plasma concentrations of dopamine and serotonine. While some metabolites were similarly affected by myo-inositol and phytase supplementation, others were distinctly differently affected. We conclude that myo-inositol, either as a directly added supplement or indirectly released from phytate upon phytase supplementation, can affect specific metabolic pathways. Additional effects found on phytase supplementation may be related to intermediary phytate degradation products. Results are indicative for innovative hypothesis to be tested in future experiments, for instance, with regard to relationships between phytase or myo-inositol supplements and bird immunity or behaviour.